Monoterpene Biosynthesis Research Articles

Identification of key gene networks controlling monoterpene biosynthesis during grape ripening by integrating transcriptome and metabolite profiling

Monoterpenes are typical aroma components of muscat grape cultivars, providing pleasant floral and fruity aromas to grapes and wines. However, the molecular mechanism of monoterpene biosynthesis between muscat and non-muscat grape remains unclear. Here, the muscat grape cultivar ‘Jumeigui’ and the non-muscat grape cultivar ‘Kyoho’ were chosen as plant materials for a comprehensive transcriptome and metabolite analysis. The gas chromatography–mass spectrometry (GC–MS) analysis demonstrated that a total of 27 and 23 monoterpene compounds were identified and quantified in the ‘Jumeigui’ and ‘Kyoho’ grape, respectively. ‘Jumeigui’ grape accumulated significantly higher concentrations of monoterpenes than ‘Kyoho’ grape. Furthermore, geraniol, linalool, geranic acid, and β-citronellol might be important odorants contributing to the floral character of the ‘Jumeigui’ grape due to the high levels odor activity values (OAVs). Transcriptome analysis demonstrated that the expression profiles of VvDXS, VvGGPPS.SSU1, VvTPS-b/g showed a positive correlation with monoterpene accumulation in grapes. In addition, the expression patterns of the genes involved in jasmonic acid (JA) synthesis and signal were also positively correlated with monoterpene accumulation. All these results will help guide the functional verification of candidate genes related to monoterpene biosynthesis, as well as identify the master transcriptional and hormonal regulators of this pathway in grapes.

Grapevine genome analysis demonstrates the role of gene copy number variation in the formation of monoterpenes.

Volatile organic compounds such as terpenes influence the quality parameters of grapevine through their contribution to the flavour and aroma profile of berries. Biosynthesis of volatile organic compounds in grapevine is relatively complex and controlled by multiple genes, the majority of which are unknown or uncharacterised. To identify the genomic regions that associate with modulation of these compounds in grapevine berries, volatile metabolic data generated via GC-MS from a grapevine mapping population was used to identify quantitative trait loci (QTLs). Several significant QTLs were associated with terpenes, and candidate genes were proposed for sesquiterpene and monoterpene biosynthesis. For monoterpenes, loci on chromosomes 12 and 13 were shown to be associated with geraniol and cyclic monoterpene accumulation, respectively. The locus on chromosome 12 was shown to contain a geraniol synthase gene (VvGer), while the locus on chromosome 13 contained an α-terpineol synthase gene (VvTer). Molecular and genomic investigation of VvGer and VvTer revealed that these genes were found in tandemly duplicated clusters, displaying high levels of hemizygosity. Gene copy number analysis further showed that not only did VvTer and VvGer copy numbers vary within the mapping population, but also across recently sequenced Vitis cultivars. Significantly, VvTer copy number correlated with both VvTer gene expression and cyclic monoterpene accumulation in the mapping population. A hypothesis for a hyper-functional VvTer allele linked to increased gene copy number in the mapping population is presented and can potentially lead to selection of cultivars with modulated terpene profiles. The study highlights the impact of VvTPS gene duplication and copy number variation on terpene accumulation in grapevine.

PbABCG1 and PbABCG2 transporters are required for the emission of floral monoterpenes in Phalaenopsis bellina.

Terrestrial plants emit volatiles into the atmosphere to attract both pollinators and the enemies of herbivores, for defense. Phalaenopsis bellina is a scented orchid species in which the main scent components are monoterpenes, including linalool and geraniol, and their derivatives. Here, we investigated whether ABC transporters are involved in floral scent emission. We carried out whole-genome identification of ABC transporter-related genes using four floral transcriptomics libraries of P. bellina. We identified 86 ABC subfamily G genes related to terpenoid transport. After comparing the gene expression patterns of P. bellina with that of Phalaenopsis aphrodite subsp. formosana, a scentless species, followed by gene-to-gene correlation analysis, PbABCG1 and PbABCG2 were selected. The temporal expression of both PbABCG1 and PbABCG2 was highly correlated with that of the key enzyme PbGDPS and the major transcription factor PbbHLH4 in monoterpene biosynthesis, with optimal expression on day5 post-anthesis. Spatial gene expression analysis showed that PbABCG1 was highly expressed in sepals, whereas PbABCG2 was expressed in the lip. Subcellular localization with a GFP fusion protein revealed that both PbABCG1 and PbABCG2 are cytoplasmic membrane proteins. Co-downregulation of PbABCG1 and PbABCG2 using both double-strand RNA interference and tobacco rattle virus-based gene silencing led to a significant decrease in monoterpene emission, accompanied by an increase in the internal monoterpene pools. Furthermore, ectopic expression of PbABCG1 and PbABCG2 in an ABC16- mutant yeast strain rescued its tolerance to geraniol. Altogether, our results indicate that PbABCG1 and PbABCG2 play substantial roles in monoterpene transport/emission in P. bellina floral scent.

LiMYB108 is involved in floral monoterpene biosynthesis induced by light intensity in Lilium 'Siberia'.

We find that the MYB family transcription factor, LiMYB108, has a novel function to regulate the floral fragrance affected by light intensity. Floral fragrance determines the commercial value of flowers and is influenced by many environmental factors, especially light intensity. However, the mechanism by which light intensity affects the release of floral fragrance is unclear. Here, we isolated an R2R3-type MYB transcription factor LiMYB108, the expression of which was induced by light intensity and located in the nucleus. Light of 200 and 600μmolm-1s-1 significantly increased the expression of LiMYB108, which was consistent with the improving trend of monoterpene synthesis under light. Virus-induced gene silencing (VIGS) of LiMYB108 in Lilium not only significantly inhibited the synthesis of ocimene and linalool, but also decreased the expression of LoTPS1; however, transient overexpression of LiMYB108 exerted opposite effects. Furthermore, yeast one-hybrid assays, dual-luciferase assays, and electrophoretic mobility shift assays (EMSA) demonstrated that LiMYB108 directly activated the expression of LoTPS1 by binding to the MYB binding site (MBS) (CAGTTG). Our findings demonstrate that light intensity triggered the high expression of LiMYB108, and then LiMYB108 as a transcription factor to activate the expression of LoTPS1, thus promoting the synthesis of the ocimene and linalool, which are important components of floral fragrance. These results provide new insights into the effects of light intensity on floral fragrance synthesis.

LcERF134 increases the production of monoterpenes by activating the terpene biosynthesis pathway in Litsea cubeba

Litsea cubeba, an aromatic species of the Lauraceae family, produces a diverse array of monoterpenes. The biosynthesis of monoterpenes is regulated by transcriptional factors (TFs), such as APETALA2/ethylene response factor (AP2/ERF). However, the regulatory mechanisms that control the AP2/ERF gene responsible for the biosynthesis of monoterpenes in L. cubeba have yet to be elucidated. Here, we identified an AP2/ERF gene, LcERF134, as an activator for the accumulation of citral and other monoterpenes. The expression level of LcERF134 was consistent with terpene synthase LcTPS42 in the pericarp. The transient overexpression of LcERF134 significantly increased monoterpene production in L. cubeba as well as the expression of rate-limiting genes involved in the monoterpene biosynthesis pathway. Furthermore, yeast one-hybrid, dual-luciferase and electrophoretic mobility shift assays demonstrated that LcERF134 activated the monoterpene biosynthesis pathway by directly binding to the GCC-box elements of the LcTPS42 and LcGPPS.SSU1 promoters. However, the overexpression of LcERF134 in tomatoes had no impact on the synthesis of monoterpenes, thus indicating that LcERF134 is a species-specific TF. Our research demonstrated that LcERF134 significantly increased the biosynthesis of monoterpenes by inducing the expression of LcTPS42 and LcGPPS.SSU1, thus offering insight into how to enhance the flavor of L. cubeba essential oil.

Genome-wide analyzation and functional characterization on the TPS family provide insight into the biosynthesis of mono-terpenes in the camphor tree

Terpene synthase (TPS) plays an important role in terpenoids biosynthesis. Cinnamomum camphora (camphor tree) contains dozens of terpenoids with medicinal value, especially borneol, which has been widely used since ancient times. However, limited information is available regarding the genome-wide identification and characterization of the TPS family in the C. camphora. In this study, 82 CcTPS genes were identified from the camphor tree genome (CTG). Gene cluster and sequence syntenic analysis suggested that tandem duplication occurred within the TPS family of the CTG, especially for the TPS-b subfamily. The chemotype-specific gene expression analysis showed significantly differential expression patterns among six chemotypes. It is worth noting that three genes (CcTPS26, CcTPS49 and CcTPS72) exhibited relatively high expression in the borneol-type camphor tree, compared to the other five chemotypes. Further functional characterization of them indicated that they were all bornyl diphosphate synthases (BPPSs), which function in catalyzing GPP into BPP and then undergoes dephosphorylation to yield borneol. This is the first report that multiple BPPSs exist within a single species. Intriguingly, CcTPS49 and CcTPS72 lead to the generation of dextral-borneol, while CcTPS26 contributes to the biosynthesis of levo-borneol. In addition, the functional characterization of another six CcTPSs suggested that they are responsible for the biosynthesis of linalool, eucalyptol and several other monoterpenes in camphor tree. In conclusion, these novel results provide a foundation for further exploration of the role of the CcTPS gene family and shed light on a better understanding of the biosynthesis and accumulation of monoterpenes in camphor tree.

Linking Pedobacter lusitanus NL19 volatile exometabolome with growth medium composition: what can we learn using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry?

Microbial metabolomics allows understanding and to comprehensively analyse metabolites, and their related cellular and metabolic processes, that are produced and released to the extracellular environment under specific conditions. In that regard, the main objective of this research is to understand the impact of culture media changes in the metabolic profile of Pedobacter lusitanus NL19 (NL19) and Pedobacter himalayensis MTCC 6384 (MTCC6384) and respective influence on the production of biotechnologically relevant compounds. Solid-phase microextraction combined with comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry with time-of-flight analyser (GC × GC-ToFMS) was applied to comprehensively study the metabolites produced by NL19 and MTCC6384 both in tryptic soy broth 100% (TSB100) and tryptic soy broth with 25% casein peptone (PC25). A total of 320 metabolites were putatively identified, which belong to different chemical families: alcohols, aldehydes, esters, ethers, hydrocarbons, ketones, nitrogen compounds, sulphur compounds, monoterpenes, and sesquiterpenes. Metabolites that were statistically different from the control (sterile medium) were selected allowing for the construction of the metabolic profile of both strains. A set of 80 metabolites was tentatively associated to the metabolic pathways such as the metabolism of fatty acids, branched-chain aminoacids, phenylalanine, methionine, aromatic compounds, and monoterpene and sesquiterpene biosynthesis. This study allowed to better understand how slight changes of the culture media and thus the composition of nutrients impair the metabolic profile of bacteria, which may be further explored for metabolomics pipeline construction or biotechnological applications.

Full-length transcriptome sequencing reveals the molecular mechanism of monoterpene and sesquiterpene biosynthesis in Cinnamomum burmannii.

Essential oil of Cinnamomum burmannii is rich in monoterpenes and sesquiterpenes and is widely used in cosmetics and medicines. Knowledge about the enzymes that catalyze the formation of monoterpenes and sesquiterpenes in C. burmannii is insufficient. Therefore, anatomy observation of C. burmannii at the four developmental stages (7days, CBS1; 14days, CBS2; 21days, CBS3, and 28days, CBS4) were conducted to elucidate the origins of essential oil production. Twelve full-length transcriptomes of C. burmannii leaves at the four stages were generated using Oxford Nanopore Technologies. GC-MS analysis revealed 15 monoterpene and sesquiterpenes dramatically increased from CBS1 to CBS4. A weighted correlation network analysis (WGCNA) in association and differentially expressed genes across four developmental stages were performed. A total of 44 differentially expressed genes (DEGs) were involved in terpenoid syntheses during leaf development. Among them, the DEGs of the mevalonate acid (MVA) pathway were predominantly expressed at CBS1, while those of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway showed increased expression from CBS2 to CBS4. Besides, fourteen genes were associated with monoterpene synthesis and nine with sesquiterpene synthesis. Functions of these DEGs were further predicted with regard to gene expression profile and phylogenetic relationship with those characterized in previous studies. In addition, 922 long noncoding RNAs (lncRNAs) were detected, of which twelve were predicted to regulate monoterpene and sesquiterpene biosynthesis. The present study provided new insights the molecular mechanisms of monoterpenoid and sesquiterpenoid syntheses of C. burmannii.

Overexpression of <i>1-deoxy-D-xylulose-5-phosphate reductoisomerase</i> enhances the monoterpene content in <i>Litsea cubeba</i>

Monoterpenes are important components of plant essential oils and have long been used as raw materials for spices and food flavorings. <italic>Litsea cubeba</italic> is an economically aromatic plant species, the fruits of which produce essential oil with monoterpenes as the dominant components. As a branch point of carbon flow in the methyl erythritol phosphate (MEP) biosynthesis pathway, 1-deoxy-D-xylo-5-phosphate reductoisomerase (DXR) is a key rate-limiting enzyme that catalyzes the MEP pathway’s second committed step. Therefore, <italic>DXR</italic> has become an effective regulation target to improve the biosynthesis of plant monoterpenes. In this study, we identified a <italic>DXR</italic> from <italic>L. cubeba</italic>, which was highly expressed in fruits and induced by darkness and MeJA. An enzyme assay showed that recombination LcDXR protein catalyzed DXP with NADPH as the cofactor. Transient overexpression of <italic>LcDXR</italic> significantly increased the content of monoterpenes in <italic>L. cubeba</italic>. Furthermore, <italic>LcDXR</italic>-overexpressing tobaccos were conducted and showed almost 5.9-fold increase in monoterpenes production, including limonene, α-pinene, eucalyptol, linalool, terpineol and camphor. Overexpression of <italic>LcDXR</italic> activated the metabolic flux of monoterpene biosynthesis through crosstalk and feedback mechanism. In addition, <italic>LcDXR</italic>-overexpressing tobaccos had no effect on phenotype of transgenic tobaccos. Our results demonstrate that <italic>LcDXR</italic> is a critical regulator of the monoterpene production in <italic>L. cubeba</italic> and other plants.

Transcriptomic and metabolic analyses reveal differences in monoterpene profiles and the underlying molecular mechanisms in six grape varieties with different flavors

Monoterpenes are important compounds determining grape floral and fruity aromas. In this study, free and bound monoterpene contents and the transcriptome profiles of six grape varieties with different flavors were determined using HS-SPME/GC-MS and RNA-sequencing technologies. The different aroma types of grape varieties had significantly different free and bound monoterpene profiles. Linalool, trans-β-ocimene, β-cis-ocimene, geraniol, and β-myrcene were the main free monoterpenes in Muscat-type grapes. In contrast, trans-β-ocimene, γ-terpinene, and β-citronellol were the main free monoterpenes in strawberry-type grapes, while limonene and β-myrcene were the main free monoterpenes in neutral grapes. The total odor activity values (OAVs) of the free monoterpenes with OAVs >1 were more than 10-times higher in Muscat-type grapes than in strawberry-type and neutral grapes, which explains the high intensity of floral, rose and sweet aromas of Muscat-type grapes. A total of 21 genes related to monoterpene biosynthesis were detected in the three types of grape varieties, among which the expression of DXS, CMK, HDS, HDR, GPPS, HMGS, HMGR, MPDC, TPS14, and GT14 was positively correlated with monoterpene accumulation, indicating these genes may contribute to the differential accumulation of monoterpene compounds in analyzed grape varieties. These results clarify the monoterpene accumulation in grapes and underlying molecular mechanism.

Engineering a universal and efficient platform for terpenoid synthesis in yeast

Engineering microbes for the production of valuable natural products is often hindered by the regulation of native competing metabolic networks in host. This is particularly evident in the case of terpenoid synthesis in yeast, where the canonical terpenoid precursors are tightly coupled to the biosynthesis of sterols essential for yeast viability. One way to circumvent this limitation is by engineering product pathways less connected to the host native metabolism. Here, we introduce a two-step isopentenol utilization pathway (IUP) in Saccharomyces cerevisiae to augment the native mevalonate pathway by providing a shortcut to the synthesis of the common terpenoid precursors, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). As such, the IUP was capable of elevating the IPP/DMAPP pool by 147-fold compared with the native pathway. We further demonstrate that cofeeding isoprenol and prenol enhances geranyl diphosphate (GPP) content for monoterpene biosynthesis. More importantly, we established a synthetic three-step route for efficient synthesis of di-and tetraterpene precursor geranylgeranyl diphosphate (GGPP), circumventing the competition with farnesyl diphosphate (FPP) for sterol biosynthesis and elevating the GGPP level by 374-fold. We combine these IUP-supported precursor-forming platforms with downstream terpene synthases to harness their potential and improve the production of industrially relevant terpenoids by several fold. Our exploration provides a universal and effective platform for supporting terpenoid synthesis in yeast.

Comprehensive identification of bHLH transcription factors in Litsea cubeba reveals candidate gene involved in the monoterpene biosynthesis pathway.

Litsea cubeba (Lour.) Person, an economically important aromatic plant producing essential oils, has lemon-like fragrance and 96.44-98.44% monoterpene contents. bHLH transcription factor plays an important role in plant secondary metabolism and terpene biosynthesis. In this study, we used bioinformatics to identify bHLH transcription factors in L. cubeba, 173 bHLH genes were identified from L. cubeba and divided these into 26 subfamilies based on phylogenetic analysis. The majority of bHLHs in each subfamily shared comparable structures and motifs. While LcbHLHs were unevenly distributed across 12 chromosomes, 10 tandem repeats were discovered. Expression profiles of bHLH genes in different tissues demonstrated that LcbHLH78 is a potential candidate gene for regulating monoterpene biosynthesis. LcbHLH78 and the terpene synthase LcTPS42 showed comparable expression patterns in various tissues and fruit development stages of L. cubeba. Subcellular localization analysis revealed that LcbHLH78 protein localizes to the nucleus, consistent with a transcription factor function. Importantly, transient overexpression of LcbHLH78 increased geraniol and linalol contents. Our research demonstrates that LcbHLH78 enhances terpenoid biosynthesis. This finding will be beneficial for improving the quality of L. cubeba and provides helpful insights for further research into the control mechanism of LcbHLH genes over terpenoid biosynthesis.

Biosynthesis, natural distribution, and biological activities of acyclic monoterpenes and their derivatives

Biosynthesis, natural distribution, and biological activities of acyclic monoterpenes and their derivatives

Cloning of StHD1 and StHD8 from Schizonepeta tenuifolia and function of regulating glandular trichome development

Hd-Zip, a unique transcription factor in plant kingdom, influences the growth, development, and secondary metabolism of plants. Hd-zip Ⅳ is thought to play an important role in trichome development of Schizonepeta tenuifolia. This study aims to explore the functions of StHD1 and StHD8 in Hd-zip Ⅳ subfamily in peltate glandular trichome development. To be specific, the expression patterns of the two genes and interaction between the proteins encoded by them were analyzed based on transcriptome sequencing and two-hybrid screening. The subcellular localization was performed and functions of the genes were verified in tobacco and S. tenuifolia. The results showed that StHD1 and StHD8 had high similarity to HD-Zip Ⅳ proteins of other plants and they all had the characteristic conserved domains of HD-Zip Ⅳ subfamily. They were located in the nucleus. The two genes mainly expressed in young tissues and spikes, and StHD1 and StHD8 proteins interacted with each other. The density and length of glandular trichomes increased significantly in tobacco plants with the overexpression of StHD1 and StHD8. Inhibiting the expression of StHD1 and StHD8 by VIGS(virus-induced gene silencing) in S. tenuifolia resulted in the reduction in the density of peltate glandular trichomes, the expression of key genes related to mono-terpene synthesis, and the relative content of limonene and pulegone, the main components of monoterpene. These results suggested that StHD1 and StHD8 of S. tenuifolia formed a complex to regulate glandular trichomes and affect the biosynthesis of monoterpenes.

Methyl Jasmonate Induces Genes Involved in Linalool Accumulation and Increases the Content of Phenolics in Two Iranian Coriander (Coriandrum sativum L.) Ecotypes.

The medicinal herb coriander (Coriandrum sativum L.), with a high linalool (LIN) content, is widely recognized for its therapeutic benefits. As a novel report, the goals of this study were to determine how methyl jasmonate (MeJA) affects total phenolic content (TPC), LIN content, flavonoid content (TFC), and changes in gene expression involved in the linalool biosynthesis pathway (CsγTRPS and CsLINS). Our findings showed that, in comparison to the control samples, MeJA treatment substantially enhanced the TPC, LIN, and TFC content in both ecotypes. Additionally, for both Iranian coriander ecotypes, treatment-induced increases in CsγTRPS and CsLINS expression were connected to LIN accumulation in all treatments. A 24 h treatment with 150 µM MeJA substantially increased the LIN content in the Mashhad and Zanjan ecotypes, which was between 1.48 and 1.69 times greater than that in untreated plants, according to gas chromatography–mass spectrometry (GC-MS) analysis. Our findings demonstrated that MeJA significantly affects the accumulation of LIN, TPC, and TFC in Iranian C. sativum treated with MeJA, which is likely the consequence of gene activation from the monoterpene biosynthesis pathway. Our discoveries have improved the understanding of the molecular mechanisms behind LIN synthesis in coriander plants.

Identification and characterization of a novel gene involved in glandular trichome development in Nepeta tenuifolia.

Nepeta tenuifolia is a medicinal plant rich in terpenoids and flavonoids with antiviral, immunoregulatory, and anti-inflammatory activities. The peltate glandular trichome (PGT) is a multicellular structure considered to be the primary storage organ for monoterpenes; it may serve as an ideal model for studying cell differentiation and the development of glandular trichomes (GTs). The genes that regulate the development of GTs have not yet been well studied. In this study, we identified NtMIXTA1, a GT development-associated gene from the R2R3 MYB SBG9 family. NtMIXTA1 overexpression in tobacco resulted in the production of longer and denser GTs. Virus-induced gene silencing of NtMIXTA1 resulted in lower PGT density, a significant reduction in monoterpene concentration, and the decreased expression of genes related to monoterpene biosynthesis. Comparative transcriptome and widely targeted metabolic analyses revealed that silencing NtMIXTA1 significantly influenced the expression of genes, and the production of metabolites involved in the biosynthesis of terpenoids, flavonoids, and lipids. This study provides a solid foundation describing a mechanism underlying the regulation of GT development. In addition, this study further deepens our understanding of the regulatory networks involved in GT development and GT development-associated metabolite flux, as well as provides valuable reference data for studying plants with a high medicinal value without genetic transformation.

Determinants of Selectivity for the Formation of Monocyclic and Bicyclic Products in Monoterpene Synthases

Monoterpene synthases (MTSs) catalyze the initial committed step in the biosynthesis of monoterpenes. MTSs control challenging reactions that involve highly reactive carbocations through exquisite steric and electrostatic confinement, in some cases with remarkable product specificity and enantioselectivity. Using two well-characterized MTSs as models, (4S)-(−)-limonene synthase (LMNS) and (+)-bornyl diphosphate synthase (BPPS), we implemented an iterative approach that involves comparative atomistic simulations and experimental testing of wild-type enzymes and thirty-six variants to identify the mechanistic underpinnings of selectivity. Free energy simulations indicate that a common reaction intermediate, the α-terpinyl cation (ATC), preferentially adopts one of two different conformations in LMNS and BPPS, thus leading to the formation of monocyclic monoterpenes in the former and bicyclic products in the latter. An assessment of the ATC binding free energy in LMNS, BPPS, and variants revealed that nonbonded interactions with active site residues explain the propensity of the ATC to assume a favored conformation that is consistent with the experimentally determined reaction outcome. The free energy of the ATC in different environments (active sites of LMNS, BPPS, and variants) correlates strongly with the ratio of monocyclic to bicyclic products in model MTSs.

Molecular evolutionary trends and biosynthesis pathways in the Oribatida revealed by the genome of Archegozetes longisetosus

Oribatid mites are a specious order of microarthropods within the Chelicerata, compromising about 11,000 described species. They are ubiquitously distributed across different microhabitats in all terrestrial ecosystems around the world and were among the first animals colonizing terrestrial habitats as decomposers and scavengers. Despite their species richness and ecological importance genomic resources are lacking for oribatids. Here, we present a 143-Mb chromosome-level genome assembly of the clonal, all-female oribatid mite species Archegozetes longisetosus Aoki, a model species used by numerous laboratories for the past 30 years. Comparative genomic and transcriptional analyses revealed patterns of reduced body segmentation and loss of the segmental identity gene abd-A within Acariformes, and unexpected expression of key eye development genes in these eyeless mites across developmental stages. Consistent with their soil dwelling lifestyle, investigation of sensory genes revealed a species-specific expansion of gustatory receptors, and evidence of horizontally transferred genes related to cell wall degradation of plant and fungal matter, both components of Archegozetes’ diet. Oribatid mites are also noted for their biosynthesis capacities and biochemical diversity. Using biochemical and genomic data, we were able to delineate the backbone biosynthesis of monoterpenes, an important class of compounds found in the major exocrine gland system of Oribatida – the oil glands. Given the mite’s strength as an experimental model, the new high-quality resources provided here will serve as the foundation for molecular research in Oribatida and will enable a broader understanding of chelicerate evolution.

Dynamic Tissue-Specific Transcriptome Changes in Response to Verticillium dahliae in Wild Mint Species Mentha longifolia.

Mentha longifolia is a wild mint species being used as a model to study the genetics of resistance to the fungal wilt pathogen Verticillium dahliae. We used high-throughput Illumina sequencing to study gene expression in response to V. dahliae inoculation in two M. longifolia USDA accessions with contrasting phenotypes: wilt-resistant CMEN 585 and wilt-susceptible CMEN 584. Roots and stems were sampled at two early post-inoculation time points, four hours and twenty-four hours, and again at ten days and twenty days post-inoculation. Overall, many more genes were differentially-regulated in wilt-resistant CMEN 585 than in wilt-susceptible CMEN 584. The greatest numbers of differentially expressed genes were found in the roots of CMEN 585 at the early time points. Specific genes exhibiting early, strong upregulation in roots of CMEN 585 but not in CMEN 584 included homologs of known plant defense response genes as well as genes involved in monoterpene biosynthesis. These genes were also upregulated in stems at the later time points. This study provides a comprehensive view of transcription reprogramming in Verticillium wilt-resistant mint, which will be the basis for further study and for molecular marker development.

SMRT sequencing of full-length transcriptome and gene expression analysis in two chemical types of Pogostemon cablin (Blanco) Benth.

BackgroundPogostemon cablin (Blanco) Benth. also called patchouli, is a traditional medicinal and aromatic plant that grows mainly in Southeast Asia and China. In China, P. cablin is divided into two chemical types: the patchouliol-type and the pogostone-type. Patchouliol-type patchouli usually grow taller, with thicker stems and bigger leaves, and produce more aromatic oil.MethodsTo better understand the genetic differences between the two chemical types that contribute to their differences in morphology and biosynthetic capabilities, we constructed de novo transcriptomes from both chemical types using the Pacific Biosciences (PacBio) Sequel platform and performed differential expression analysis of multiple tissues using Illumina short reads.ResultsIn this study, using single-molecule real-time (SMRT) long-read sequencing, we obtained 22.07 GB of clean data and 134,647 nonredundant transcripts from two chemical types. Additionally, we identified 126,576 open reading frames (ORFs), 100,638 coding sequences (CDSs), 4,106 long noncoding RNAs (lncRNAs) and 6,829 transcription factors (TFs) from two chemical types of P. cablin. We adopted PacBio and Illumina sequencing to identify differentially expressed transcripts (DEGs) in three tissues of the two chemical types. More DEGs were observed in comparisons of different tissues collected from the same chemical type relative to comparisons of the same tissue collected from different chemical types. Furthormore, using KEGG enrichment analysis of DEGs, we found that the most enriched biosynthetic pathways of secondary metabolites of the two chemical types were “terpenoid backbone biosynthesis”, “phenylpropanoid biosynthesis”, “plant hormone signal transduction”, “sesquiterpenoid and triterpenoid biosynthesis”, “ubiquinone and other terpenoid-quinone biosynthesis”, “flavonoid biosynthesis”, and “flavone and flavonol biosynthesis”. However, the main pathways of the patchouliol-type also included “diterpene biosynthesis” and “monoterpene biosynthesis”. Additionally, by comparing the expression levels of the three tissues verified by qRT-PCR, more DEGs in the roots were upregulated in the mevalonate (MVA) pathway in the cytoplasm, but more DEGs in the leaves were upregulated in the methylerythritol phosphate (MEP) pathway in the plastid, both of which are important pathways for terpenoids biosynthesis. These findings promote the study of further genome annotation and transcriptome research in P. cablin.